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Recombinant Rabbit Monoclonal
Anti-Di-Methyl-Histone H3 (Lys122) Rabbit mAb
Catalog Number: PTM-645RM
$ 385

Clone Number: 76-H1L3

Host: Rabbit Clonality: Recombinant Monoclonal

Applications: WB ICC/IF IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K122me2

Product Size
100 μl
Quantity

Shipping: Ambient temperature

Order online or send purchase order to info@ptmbio.com

FAQ Technical Support Protocols

General Information
Isotype IgG
Conjugate Unconjugated
Synonyms H3K122me2
UniProt ID

/

Immunogen
MW (kDa) 15
Specificity Anti-Dimethyl-Histone H3 (Lys122) Rabbit mAb detects histone H3 only when it is Dimethylated at Lys122.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:1000 Human, Mouse, Rat
ICC/IF 1:50 Human
IP 1:25-1:100 Human
ChIP 6 ug/ 5×106 cells Human
Properties
Purity Protein G and immunogen affinity purified
Constituents PBS, Glycerol, BSA
Storage Store at -20°C. Avoid freeze/thaw cycles.
Stability Stable for 12 months from date of receipt/reconstitution.
Target Information

Background

Post translational modifications on histones include acetylation, methylation, phosphorylation and some new acylation modifications found in recent years. These histone modifications directly affect the binding of chromatin to transcription factors or other epigenetic regulators, and change genome stability and gene transcription. Histone methylation usually occurs in lysine and arginine residues of core histones. Histone methylation can promote or inhibit gene transcription, depending on whether histone methylation occurs on lysine or arginine and the number of methylation groups (lysine can be monomethyl, dimethyl and trimethylated, arginine can be monomethyl, symmetric and asymmetric dimethyl). Histone lysine methylation usually occurs on lysine 4, 9, 27, 36, 79 of histone H3 and lysine 20 of histone H4; Arginine methylation usually occurs on arginine 2, 8, 17, 26 of histone H3 and arginine 3 of histone H4. Protein methylase (HMT) and histone demethylase (HDM) are the main regulators of methylation modification. H3K122me2 is a newly discovered modification site.

Cellular location

Nucleus

Images
Dot Blot

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Immunogen peptide quantity: 1: 1ng, 4 ng, 16 ng, 64 ng, 2-4: 64 ng
Exposure time: 60 seconds
The list of peptides is included in the table below.

WB

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab:1: HeLa cytosol, 2: HeLa nuclear
Protein loading quantity: 20 μg
Exposure time: 60 seconds
Predicted band size: 15 KDa
Observed band size: 15 KDa

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab:1: Mouse brain, 2: Rat brain, 3: Recombinant histone H3
Protein loading quantity: 20 μg
Exposure time: 60 seconds
Predicted band size: 15 KDa
Observed band size: 15 KD

IP

IP of HeLa nuclear extracts
IP ab incubation condition: PTM-645RM, 4°C
overnight, 1:100 dilution
WB primary ab incubation condition: PTM645RM, room temperature 2h, 1:1000 dilution
Secondary ab: Anti-Rabbit IgG for IP (HRP)
Blocking buffer and concentration: 5%
NFDM/TBST
Diluting buffer and concentration: 5%
NFDM/TBST
1: 5% Input
2: IP with PTM-645RM
3: IP with Rabbit monoclonal IgG Isotype
Observed MW: 15 kDa
Exposure time: 120 s

ICC/IF

Cell line: HeLa
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% TritonX-100
Primary ab dilution: 1:50
Primary incubation condition: 4℃ overnight
Secondary ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (Blue)
Counter stain: Tubulin (Red)
Comment: Color green is the positive signal for
PTM-645RM

ChIP

Cell type: HeLa
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5×106
cells
Amount of Ab per IP: 6 μg
Beads type and amount per IP: 50 μL of Protein A MagBeads
Comment: The ChIP was performed with 6 μg of normal Rabbit IgG as a negative control. Real
time quantitative PCR was performed on immunoprecipitated DNA using primers specific for the
human GAPDH CDS region, RPL30, LDHA CDS region, FOXO3a-promoter, FOXO3adownstream, RAB20 and TuBBP10. Data are presented as enrichment of each sample relative
to total amount of input chromatin at each amplicon.

Research Use

For research use only, not for use in diagnostic procedures.